To be determined.
The present invention relates to the field of bone morphogenetic proteins and more particularly to genes and proteins in the BMP-1/Tld family.
Bone formation in mammals such as mice and humans is governed by a set of bone morphogenetic proteins (BMP). Of the seven BMPs known to participate in osteogenesis, six (designated BMP-2 through BMP-7) belong to the TGF-xcex2 super family. The seventh BMP (designated BMP-1) is not TGF-xcex2-like, but instead appears to derive from a different gene family. The BMP-1 gene family members typically contain the following domains: an astacin-like metalloprotease domain, one or more EGF-like motifs which in other proteins are thought to bind Ca++, and a number of CUB domains. A CUB domain is a motif that mediates protein-protein interactions in complement components C1r/C1s which has also been identified in various proteins involved in developmental processes. BMP-1 was described, at the nucleotide sequence level, by Wozney, J. M., et al., Science 242:1528-1534 (1988).
The mammalian BMP-1 domain structure is shared by proteins found in other non-mammalian species. These proteins include Drosophila tolloid (Tld) (Shimell, M. J., Cell 67:469-481 (1991)), a tolloid-like Drosophila gene product (Tlr-1 or tolkin) (Nguyen, T., Dev. Biol. 166:569-586 (1994) and Finelli, A. L., et al., Genetics 141:271-281 (1995)), a sea urchin BMP-1 homolog (suBMP-1) (Hwang, S. P., et al., Development 120:559-568 (1994)), two related sea urchin developmental gene products, SpAN and BP10 (Reynolds, S. D., et al., Development 114:769-786 (1992) and Lepage, T., et al., Development 114:147-164 (1992)), a Xenopus BMP-1 (xBMP-1) (Maeno, M. et al., Gene 134:257-261 (1993), a Xenopus tolloid (Lin. J., et al., Develop. Growth Differ. 39:43-51 (1997), a tolloid-like Xenopus gene product named xolloid (Piccolo, S. et al., Cell 91:407-416 (1997), a related member of the family isolated from zebrafish and called zebrafish tolloid (Bladder, P. et al., Science 278:1937-1940 (1997), a mammalian tolloid (mTld) (Takahara, K. et al., J. Biol. Chem. 269:32572-32578 (1994)) and a mammalian tolloid-like gene (mTll-1) (Takahara, K. et al., Genomics 34:157-165 (1996)). Some of the nucleic acid sequences of the genes that encode these proteins are known. The mammalian BMP1 gene encodes both the BMP-1 protein and the mTld protein, albeit on two distinct, alternately spliced mRNA molecules. The papers mentioned in this paragraph are incorporated herein by reference.
The present invention is summarized in that a novel mammalian tolloid-like gene product (mTll-2) and its cognate polynucleotide gene, mTLL-2, are distinct from mTld and from all other known BMP-1-related gene products and their cognate polynucleotides, including mTll (now designated mTll-1). The human and murine versions of the gene are reported herein as SEQ ID NO:1 and SEQ ID NO:3, respectively. The amino acid sequences encoded by each are presented as SEQ ID NO:2 and SEQ ID NO:4, respectively.
It is an object of the present invention to provide a gene and gene product involved in the deposition of extracellular matrix in vertebrates (e.g., in osteogenesis).
It is another object of the present invention to provide a target molecule for rational development of a drug for inhibiting activity of the tolloid-like genes to treat fibrosis, scarring, keloids, surgical adhesions, and the like.
It is yet another object of the present invention to provide a recombinant DNA construct, and a protein encoded by the construct, for use in accelerated wound and fracture healing.
It is still another object of the present invention to provide markers that map to the 10q24 region of human chromosome 10 and to the distal end of chromosome 19 in mice.
It is still another object of the present invention to provide nucleotide sequences that function as probes for a non-BMP-1 bone morphogenetic protein gene in mammalian cells.
Other objects, features, and advantages of the present invention will become apparent upon consideration of the following detailed description considered in conjunction with the accompanying drawings.